Release of euchromatic segments by MgCa-dependent autodigestion of chromatin

Paul, Izhak J.; Duerksen, Jacob D.

doi:10.1016/0006-291x(76)90015-2

Cited by 24 publications

(2 citation statements)

References 16 publications

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“…Glycerol Gradient Centrifugation of Nuclease-treated Chromatin: Two 1 mg samples of chromatin in glycine buffer (pH 7.1) with 25 pM CaC2 were incubated with 30 units micrococcal nuclease/ml at 300, one for 5 minutes and the other for 30 minutes. Each of these samples was cooled and layered onto a 7.6-76% glycerol gradient and centrifuged for 16 hours at 20,000 rpm at 10 in a Spinco preparative ultracentrifuge as previously described 20…”

Section: Methodsmentioning

confidence: 99%

Satellite DNA sequence content of polylysine-titratable and nuclease-resistant fractions of mouse liver hepatoma chromatin

Duerksen

Paul

1976

Nucleic Acids Research

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Micrococcal nuclease digestion of mouse TLT liver hepatoma chromatin proceeds rapidly to the point where approximately 35% of the DNA is recoverable by centrifugation of the chromatin DNA through 3M CsCl. The satellite DNA sequence content of this recoverable DNA is the same as whole chromatin DNA (10%). The 11s (penultimate digestion product) monomer, as well as intermediate multiples and relatively undigested large chromatin segments are separable on steep hlycerol gradients. The DNA isolated from these fractions also contains the normal 10% satellite DNA content. Progressive polylysine titration of chromatin followed by nuclease digestion gives anomalous recoveries of DNA but, nonetheless, the satellite sequence content titration of chromatin, followed by pronase and then nuclease digestion, again gave recoverable DNA with a satellite sequence content of 10%. These results are discussed in terms of the conclusion that nucleosome (or upsilon-body) structures are distributed in a random fashion over the genome.

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Section: Methodsmentioning

confidence: 99%

Satellite DNA sequence content of polylysine-titratable and nuclease-resistant fractions of mouse liver hepatoma chromatin

Duerksen

Paul

1976

Nucleic Acids Research

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“…Toward this end, attempts have been made to separate chromatin into transcribed and nontranscribed components. Methods of fractionation include differential centrifugation (Frenster et al, 1963;Frenster, 1965; ; Yasmineh & Yunis, 1969; Yunis & Yasmineh, 1970;Duerksen & McCarthy, 1971;McCarthy et al, 1973; Murphy et al, 1973;Chesterton et al, 1974;Berkowitz & Doty, 1975; Burkholder & Weaver, 1975;Charles et al, 1975;Howk et al, 1975;Paul & Duerksen, 1976), differential solubility (Jensen & Chalkey, 1968;Marushige & Bonner, 1971; Billing & Bonner, 1972;Bonner et al, 1973;Gottesfeld et al, 1974Gottesfeld et al, ,1975Gottesfeld et al, ,1976; Arnold & Young, 1974; Pederson & Bhorjee, 1975), hydroxylapatite thermal chromatography (McConaughy & McCarthy, 1972), agarose gel exclusion chromatography (Janowski et al, 1972;McCarthy et al, 1973;Anderson et al, 1975;Pays & Flamand, 1976; Kreig & Wells, 1976), ECTHAM-cellulose ion-exchange chromatography (Reeck et al, 1972(Reeck et al, , 1974 Simpson & Reeck, 1973; ; Simpson, 1975;Howk et al, 1975;Levner et al, 1975;Keller et al, 1975) and others…”

Section: Hydroxylapatite Thermal Fractionation Of Chromatin and Dna1mentioning

confidence: 99%

Hydroxylapatite thermal fractionation of chromatin and DNA

Saffitz¹,

Caplan²

1978

Biochemistry

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Chromatin and DNA from developing muscle cultures were fractionated by hydroxylapatite thermal chromatography on the basis of differential thermal stability. A thermal chromatography system was developed in which protein mediated thermal stability of chromatin DNA was maximally expressed. The resulting chromatin and DNA elution profiles were similar to thermal denaturation profiles in low ionic strength solution. Additional studies showed this system was able to detect protein stabilization of DNA in in vitro nucleohistone preparations. Although some protein remained bound to hydroxylapatite during chromatin thermal elution, it did not affect the denaturation or elution behavior of free DNA on the same column. These studies show that fragments of chromatin or DNA can be segregated on the basis of differential thermal stability by hydroxylapatite chromatography.

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Periodicity and fragment size of DNA from mouse TLT hepatoma chromatin and chromatin fractions using endogenous and exogenous nucleases

1978

Mol Cell Biochem

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The action of micrococcal nuclease, DNase I and DNase II on mouse TLT hepatoma chromatin revealing the periodicity of its structure as visualized by denaturing and non-denaturing gel electrophoresis, was consistent with the action of these enzymes on other chromatins. Micrococcal nuclease showed a complex subnucleosome fragment pattern based on multiples of 10 base pairs with a prominant couplet at 140/160 base pairs and the absence of the 80 base pair fragment. This couplet of the core and minimal nucleosome fragments was conspicuously present in the mononucleosomes found in the 11S fractions of a glycerol gradient centrifugation. DNase I and II produced a fairly even distribution of a 10 base pair increasing series of fragments to about 180 base pairs, a pattern also repeated in the DNA of nucleosome glycerol-gradient fractions. In limited digestions by these nucleases multinucleosomic DNA fragments are pronounced. These fragment lengths are multiples of an estimated average repeat length of nucleosome DNA of 180 base pairs. The action of the endogenous Mg/Ca-stimulated endonuclease produced only limited cuts in the hepatoma chromatin resulting primarily in multi-nucleosomic DNA fragment lengths and only upon lengthy digestion limited subnucleosomic, 10-base-pair multiple fragments are produced. The putative euchromatin-enriched fractions (50-75S) of the glycerol gradient centrifugation of autodigested chromatin, similarly, contained primarily the multinucleosomic DNA fragment lengths. These results are consistent with our previous electron microscopic demonstration that autodigested chromatin as well as the putative euchromatin-enriched fractions were composed of multi-nucleosomic chromatin segments containing a full complement of histones.

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Release of euchromatic segments by MgCa-dependent autodigestion of chromatin

Cited by 24 publications

References 16 publications

Satellite DNA sequence content of polylysine-titratable and nuclease-resistant fractions of mouse liver hepatoma chromatin

Satellite DNA sequence content of polylysine-titratable and nuclease-resistant fractions of mouse liver hepatoma chromatin

Hydroxylapatite thermal fractionation of chromatin and DNA

Periodicity and fragment size of DNA from mouse TLT hepatoma chromatin and chromatin fractions using endogenous and exogenous nucleases

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